Grape molasses (GM), produced from grapes, is a traditional Iranian food and is widely consumed in Iran. However, GM adulteration is among the most widespread illegitimate procedures involving contamination of food with foreign materials, such as adding sugar-water solution, date syrup, sugar beet syrup, and grape sauce. This study used stable carbon 13C/12C isotope ratio analysis method to detect adulteration of GM samples with glucose syrups (GS), fructose syrups (FS), and beet sugar syrups (BS) at the ratio of 0%, 10%, 30%, and 50% (by weight). Physicochemical properties of GM including °Brix, conductivity, specific gravity, pH, moisture content, ash content, hydroxymethyl furfural, sugar content, and rheological properties of samples were investigated. The δ13C isotope ratio of the GM was determined as -26.61%, that of the GS as -13.23%, that of the FS as -13.42%, and that of the BS as -16.58%. The δ13C isotope ratio increased by the addition of adulterant syrups to GM. The addition of each adulterant syrup had a different effect on the physicochemical parameters; however, the °Brix and specific gravity had a positive correlation with the δ13C isotope ratio results. The magnitudes of G' and G" increase with an increase in frequency representing the viscoelastic behavior of samples. The obtained results of this study suggest the use of δ13C isotope ratio method as a fast and accurate method to investigate the adulteration of grape molasses.